Sensitization of Gram-Negative Bacteria to Aminoglycosides with 2-Aminoimidazole Adjuvants
Abstract
:1. Introduction
2. Results
2.1. Initial Screen
2.2. Follow-Up Screen of Additional Analogues
2.3. Activity of Lead Compounds with Additional Aminoglycosides
2.4. Activity of Lead Compounds against Other Gram-Negative Pathogens
2.5. Effect of Divalent Cations on Adjuvant Activity
2.6. Evolution of TOB Resistance in the Presence and Absence of Adjuvant and Identification of Mutated Genes in Evolved Strains
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains, Media, and Antimicrobial Agents
4.2. Broth Microdilution Method for Minimum Inhibitory Concentration Determination
4.3. Broth Microdilution Method for Measurement of Aminoglycoside Potentiation
4.4. Evolution of Tobramycin Resistance in A. baumannii
4.5. Sequencing Data Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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AB5075 | AB19606 | |||
---|---|---|---|---|
Compound | Concentration (μM) | TOB MIC (μg/mL) (Fold-Reduction) | Concentration (μM) | TOB MIC (μg/mL) (Fold-Reduction) |
128 | 4 | |||
1 | 30 | 2 (64) | 30 | 1 (4) |
2 | 30 | 16 (8) | 30 | 1 (4) |
3 | 15 | 64 (2) | 30 | 0.5 (8) |
4 | 60 | 32 (4) | 60 | 2 (2) |
5 | 7.5 | 32 (4) | 15 | 2 (2) |
AB5075 | AB19606 | |||
---|---|---|---|---|
Compound | Concentration (μM) | TOB MIC (μg/mL) (Fold-Reduction) | Concentration (μM) | TOB MIC (μg/mL) (Fold-Reduction) |
128 | 4 | |||
6 | 30 | 4 (32) | 30 | 0.5 (8) |
7 | 30 | 2 (64) | 15 | 0.5 (8) |
8 | 60 | 4 (32) | 60 | 2 (2) |
9 | 60 | 16 (8) | 60 | 0.0625 (64) |
10 | 30 | 32 (4) | 60 | 0.25 (16) |
11 | 60 | 16 (8) | 30 | 1 (4) |
12 | 60 | 8 (16) | 60 | 0.5 (8) |
Compound | Concentration (μM) | STM MIC (μg/mL) (Fold-Reduction) | Concentration (μM) | KAN MIC (μg/mL) (Fold-Reduction) | Concentration (μM) | NEO MIC (μg/mL) (Fold-Reduction) | Concentration (μM) | GEN MIC (μg/mL) (Fold-Reduction) |
---|---|---|---|---|---|---|---|---|
2048 | 2048 | 64 | 512 | |||||
1 | 30 | 128 (16) | 15 | 2048 (0) | 15 | 32 (2) | 15 | 32 (16) |
6 | 30 | 32 (64) | 30 | 1024 (2) | 30 | 16 (4) | 30 | >512 (0) |
7 | 15 | 128 (16) | 7.5 | 2048 (0) | 7.5 | >64 (0) | 7.5 | 256 (2) |
8 | 30 | 512 (4) | 60 | 2048 (0) | 60 | 4 (16) | 60 | 16 (32) |
Compound | Concentration (μM) | STM MIC (μg/mL) (Fold-Reduction) | Concentration (μM) | KAN MIC (μg/mL) (Fold-Reduction) | Concentration (μM) | NEO MIC (μg/mL) (Fold-Reduction) | Concentration (μM) | GEN MIC (μg/mL) (Fold-Reduction) |
---|---|---|---|---|---|---|---|---|
512 | 8 | 8 | 32 | |||||
1 | 30 | 128 (4) | 15 | >8 (0) | 15 | 4 (2) | 15 | 4 (8) |
3 | 30 | 64 (8) | 30 | 2 (4) | 30 | 1 (8) | 30 | 1 (32) |
5 | 15 | 64 (8) | 15 | 4 (2) | 15 | 2 (4) | 15 | 2 (16) |
6 | 30 | 16 (32) | 30 | 1 (8) | 30 | 1 (8) | 30 | 32 (0) |
7 | 7.5 | 512 (0) | 15 | 1 (8) | 15 | 2 (4) | 15 | 4 (8) |
10 | 60 | 16 (32) | 60 | 0.5 (16) | 60 | 0.5 (16) | 60 | 0.25 (128) |
30 | 2 (4) | 45 | 2 (16) |
Strain | TOB MIC (μg/mL) | TOB MIC (μg/mL) + 1 (μM) | TOB MIC (μg/mL) + 3 (μM) | TOB MIC (μg/mL) + 5 (μM) | TOB MIC (μg/mL) + 10 (μM) | TOB MIC (μg/mL) + 11 (μM) | TOB MIC (μg/mL) + 12 (μM) |
---|---|---|---|---|---|---|---|
KP1705 | 32 | (15) 16 | (15) 32 | (15) 16 | (60) 16 | (60) 32 | (60) 8 |
KP43816 | 1 | (15) 0.125 | (15) 0.5 | (15) 0.125 | (30) 0.25 | (60) 0.5 | (10) 0.5 |
EC197 | 32 | (3.75) 32 | (7.5) 16 | (7.5) 16 | (15) 16 | (60) 16 | (15) 16 |
EC199 | 0.5 | (7.5) 0.5 | (7.5) 0.5 | (7.5) 0.5 | (15) 0.5 | (60) 0.5 | (15) 0.5 |
AB5075 Passaged in TOB Only | AB5075 Passaged in TOB + 1 (15 μM) | |
---|---|---|
Day 0 | 128 | 128 |
Day 2 | 1024 | 256 |
Day 4 | 1024 | 1024 |
Day 6 | 4096 | 1024 |
Day 8 | 4096 | 2048 |
Gene Product (Gene) | Locus Tag | # of Mutations | Type of Mutation |
---|---|---|---|
Alkyl hydroperoxide reductase subunit F (ahpF) | A591_RS08360 | 11 | 8 snp; 3 complex |
Aldehyde dehydrogenase | A591_RS16100 | 3 | 1 snp; 2 complex |
Bacteriophage protein | A591_RS13885 | 3 | 2 snp; 1 complex |
Stress-induced protein | A591_RS20205 | 1 | snp |
Replication initiation factor domain-containing protein | A591_RS15850 | 1 | Insertion |
Ig-like domain repeat protein | A591_RS19590 | 1 | snp |
Hypothetical protein (elongation factor Tu (tuf)) | A591_RS01280 | 1 | snp |
Hypothetical protein (replication protein) | A591_RS20250 | 1 | Insertion |
Unknown hypothetical protein | A591_RS20245 | 1 | Insertion |
Unknown hypothetical protein | A591_RS15845 | 1 | Insertion |
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Crotteau, A.N.; Hubble, V.B.; Marrujo, S.A.; Mattingly, A.E.; Melander, R.J.; Melander, C. Sensitization of Gram-Negative Bacteria to Aminoglycosides with 2-Aminoimidazole Adjuvants. Antibiotics 2023, 12, 1563. https://doi.org/10.3390/antibiotics12111563
Crotteau AN, Hubble VB, Marrujo SA, Mattingly AE, Melander RJ, Melander C. Sensitization of Gram-Negative Bacteria to Aminoglycosides with 2-Aminoimidazole Adjuvants. Antibiotics. 2023; 12(11):1563. https://doi.org/10.3390/antibiotics12111563
Chicago/Turabian StyleCrotteau, Ashley N., Veronica B. Hubble, Santiana A. Marrujo, Anne E. Mattingly, Roberta J. Melander, and Christian Melander. 2023. "Sensitization of Gram-Negative Bacteria to Aminoglycosides with 2-Aminoimidazole Adjuvants" Antibiotics 12, no. 11: 1563. https://doi.org/10.3390/antibiotics12111563